Weak partitioning chromatography for anion exchange purification of monoclonal antibodies
- 26 August 2008
- journal article
- Published by Wiley in Biotechnology & Bioengineering
- Vol. 101 (3), 553-566
- https://doi.org/10.1002/bit.21923
Abstract
Weak partitioning chromatography (WPC) is an isocratic chromatographic protein separation method performed under mobile phase conditions where a significant amount of the product protein binds to the resin, well in excess of typical flowthrough operations. The more stringent load and wash conditions lead to improved removal of more tightly binding impurities, although at the cost of a reduction in step yield. The step yield can be restored by extending the column load and incorporating a short wash at the end of the load stage. The use of WPC with anion exchange resins enables a two‐column cGMP purification platform to be used for many different mAbs. The operating window for WPC can be easily established using high throughput batch‐binding screens. Under conditions that favor very strong product binding, competitive effects from product binding can give rise to a reduction in column loading capacity. Robust performance of WPC anion exchange chromatography has been demonstrated in multiple cGMP mAb purification processes. Excellent clearance of host cell proteins, leached Protein A, DNA, high molecular weight species, and model virus has been achieved. Biotechnol. Bioeng. 2008;101: 553–566.This publication has 23 references indexed in Scilit:
- High‐throughput screening of chromatographic separations: I. Method development and column modelingBiotechnology & Bioengineering, 2008
- Fragments of protein A eluted during protein A affinity chromatographyJournal of Chromatography A, 2007
- Application of a chromatography model with linear gradient elution experimental data to the rapid scale-up in ion-exchange process chromatography of proteinsJournal of Chromatography A, 2007
- An exclusion mechanism in ion exchange chromatographyBiotechnology & Bioengineering, 2006
- Comparison of the Escherichia coli proteomes for recombinant human growth hormone producing and nonproducing fermentationsProteomics, 2003
- Industrial Purification of Pharmaceutical Antibodies: Development, Operation, and Validation of Chromatography ProcessesBiotechnology and Genetic Engineering Reviews, 2001
- Use of the steric mass action model in ion-exchange chromatographic process developmentJournal of Chromatography A, 1999
- The focusing positions of polypeptides in immobilized pH gradients can be predicted from their amino acid sequencesElectrophoresis, 1993
- Étude de l'élution non-linéaire en chromatographie en phase liquide préparativeJournal of Chromatography A, 1980
- Separation of neutral proteins on ion-exchange resinsBiochemical Journal, 1955